Edwards Medium Base, Modified M748

Edwards Medium Base, Modified is a selective medium for the rapid isolation of Streptococcus agalactiae and other
streptococci associated with mastitis and also from other clinical samples.
Composition**
Ingredients Gms / Litre
Peptic digest of animal tissue 10.000
Beef extract 10.000
Esculin 1.000
Sodium chloride 5.000
Crystal violet 0.0013
Thallous sulphate 0.330
Agar 15.000
Final pH ( at 25°C) 7.4±0.2
**Formula adjusted, standardized to suit performance parameters

Directions
Suspend 41.33 grams in 1000 ml distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving
at D 115°C for 20 minutes. Cool to 50°C and aseptically add 5 to 7% v/v sterile sheep blood. Mix well and pour into sterile
Petri plates.
(D corresponds to 10lbs pressure)

Principle And Interpretation

Streptococci are gram-positive facultatively anaerobic bacteria, which constitute normal commensal flora of mouth, skin,
intestine and upper respiratory tract of humans. Group B Streptococci are an important cause of systemic infections in infants
and occasionally of bacterial endocarditis (1). Mastitis is a disease of cattle caused by the organisms Streptococcus agalactiae
. It belongs to the Lancefield group B Streptococci .
The most common selective agents used for selective isolation of Streptococci are crystal violet and thallium salts. A selective
medium containing crystal violet was used by Haxthausen to isolate skin Streptococci (2). Subsequently it was observed that
Streptococci from milk were able to grow on Gentian Violet Blood Agar whereas the other saprophytic milk bacteria were
inhibited on this medium (3). An Esculin Blood Agar containing crystal violet was used by Edwards to isolate the causative
agent of mastitis (4). A similar medium containing thallous acetate was also used to isolate the causative agent of mastitis (5).
Peptic digest of animal tissue and beef extract serve as sources of carbon, nitrogen and other essential nutrients. Esculin helps to
differentiate esculin-positive (group D Streptococci) organisms from esculin- negative ( S. agalactiae ) organisms. Sodium
chloride helps to maintain the osmotic equilibrium of the medium. Crystal violet and thallous sulphate serve as the selective
agents for Streptococci. Supplementation with blood provides additional nutrients in addition to serving as an indicator of
haemolysis. Mastitis Streptococci show alpha, beta or gamma type of haemolysis. Esculin differentiates esculin- positive group
D Streptococci (black colonies) from esculin-negative Streptococcus agalactiae (blue to colourless colonies).
Centrifuged test milk sample is directly inoculated on the surface of the medium plate. Esculin-negative (blue to colourless)
S. agalactiae organisms are further subcultured for identification tests.

Quality Control
Appearance
Cream to yellow homogeneous free flowing powder
Gelling
Firm,comparable with 1.5% Agar gel
Colour and Clarity of prepared medium

Please refer disclaimer Overleaf.

HiMedia Laboratories Technical Data

Basal medium :Amber coloured, clear to slightly opalescent gel. After addition of 5-7% v/v sterile defibrinated sheep blood :
Cherry red coloured opaque gel forms in Petri plates
Reaction
Reaction of 4.13% w/v aqueous solution at 25°C. pH : 7.4±0.2
pH
7.20-7.60
Cultural Response
Cultural characteristics observed with added 5-7%v/v sterile defibrinated sheep blood after an incubation at 35-37°C for
24-48 hours .
Cultural Response
Organism Inoculum Growth Recovery Colour of
(CFU) Colony
Cultural Response
Enterococcus faecalis ATCC 50-100 good-luxuriant >=50% black
29212
Escherichia coli ATCC >=10³ inhibited 0%
25922
Staphylococcus aureus >=10³ inhibited 0%
ATCC 25923
Streptococcus agalactiae 50-100 good-luxuriant >=50% colourless, w/
ATCC 13813 haemolysis

Storage and Shelf Life

Store below 30°C in tightly closed container and the prepared medium at 2 - 8°C. Use before expiry date on the label.

Reference
1. Cruickshank R., Duguid J. P., Marmion B. P. , Swain R. H. A., (Eds.), 1975, Medical Microbiology, The Practice of Medical
Microbiology , 12th Edition, Vol. II , Churchill Livingstone.
2. Haxsthausen H., 1927, Ann. Derm. Suph., 8.201.
3. Bryan C. S., 1932, Am. J. Public Health, 22. 749.
4. Edwards S. J., 1933, J. Comp. Path. Therap., 46:211-217.
5. McKenzie D. A., 1941, Vet. Rec., 53:473-480.

Revision : 1 / 2011

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